Introducing CellGenix T-cell medium — early onset of T-cell expansion for faster T-cell therapy manufacturing

14 Apr 2020

Written by CellGenix

CAR-T Cell therapy Manufacturing & scale-up Opinion Spotlight on allogeneic CAR-T UK and Europe

Achieve an early onset of T-cell expansion and sustained viability with the novel CellGenix (Freiburg im Breisgau, Germany) T-cell medium (CellGenix® TCM).

Serum-free T-cell therapy manufacturing

CellGenix (Freiburg im Breisgau, Germany) T-cell medium (CellGenix® TCM) offers a serum-free and xeno-free alternative for rapid expansion of functional human T-cells. Due to stable glutamine in the formulation it is ready-to-use for T-cell cultures without the need for supplementation with human serum or glutamine. Many current T-cell therapy protocols rely on the addition of human serum.

Eliminating the use of human serum will reduce the failure rate in your manufacturing process due to the high lot-to-lot inconsistencies of serum. Since human serum is a limited resource and might not be available in large quantities it is unsuitable for commercial scale manufacturing. Furthermore, the human origin of serum poses a certain risk of containing adventitious agents and therefore does not meet global regulatory guidelines.

Early onset of T-cell expansion and sustained viability

T-cell culture in CellGenix TCM results in high cell numbers early after activation and throughout cell culture with high viability. This early onset of T-cell expansion allows for faster T-cell therapy manufacturing, which can significantly reduce your cost of goods.

Figure 1: Fold T-cell expansion and cell viability on the indicated day of culture: CD3+ T-cells were activated with anti-CD3/anti-CD28-coupled beads and cultivated in CellGenix TCM or competitor media in the presence of IL-7 and IL-15 (10 ng/ml, CellGenix). On the indicated day cells were labeled with 7-AAD and viability was determined by flow cytometry. Median and spread of n=6 donors are displayed for expansion (upper panels) and cell viability (lower panels).

Promotion of a central memory and early differentiated memory T-cell phenotype

During T-cell culture in CellGenix TCM a ‘young’ phenotype of naÃ¯ve T-cells/memory stem T-cells (T_N/SCMcells) or central memory T-cells (T_CM cells) is acquired. These phenotypes represent the most undifferentiated memory T-cell phenotypes, meaning that they have the highest proliferation potential, self-renewal properties and longer survival rates.

It has been shown that the engraftment and persistence of T-cell products in patients is associated with a T_CM phenotype, rather than more differentiated memory phenotypes [1]. T_SCM are considered as even less differentiated cells with potentially even better persistence. It has been reported that for CD19 CAR-T cells the expansion of patient-infused cells is correlated with the frequency of CD8⁺ T_SCM-like cells [2]. It is therefore beneficial for your T-cell therapy product to obtain a central memory and early differentiated memory T-cell phenotype.

Figure 2: Memory phenotype of starting population and after expansion in CellGenix TCM: CD3+ T-cells were activated with anti-CD3/anti-CD28-coupled beads and cultivated in CellGenix TCM or competitor media, in the presence of interleukin (IL)-7 and IL-15 (10 ng/ml, CellGenix). On day 0 (left panel) and day 11 (right panel) the expression of memory cell markers was determined by flow cytometry for three donors. T_CM, central memory T-cells; T_N, naÃ¯ve T-cells; T_SCM, memory stem T-cells; T_EM, effector memory T-cells; T_EMRA, terminally differentiated effector T-cells.

High proportion of cytokine producing cells including polyfunctional cells

An important aspect of T-cell functionality is the secretion of cytokines such as interleukin-2 (IL-2), interferon-gamma (IFN-Î³) and tumor necrosis factor-alpha (TNF-Î±). After cultivation in CellGenix TCM a high proportion of cytokine producing cells is obtained including polyfunctional cells (expressing more than one cytokine).

A recent report suggests that the combination of cell frequency and cytokine production levels of polyfunctional CD19 CAR-T cells is correlated with the clinical outcome in patients with non-Hodgkin lymphoma [3]. Obtaining cytokine secreting T-cells, in particular polyfunctional cells, could therefore have a positive effect on the therapeutic response of your T-cell therapy product.